中山大学学报自然科学版 ›› 2018, Vol. 57 ›› Issue (1): 29-35.

• 论文 • 上一篇    下一篇

宝山堇菜Pb耐性相关基因筛选及表达特性

邓冬梅1,艾红霞2,邓金川2,廖斌2   

  1. 1. 广西科技大学生物与化学工程学院∥广西糖资源绿色加工重点实验室,广西 柳州 545006;
    2. 中山大学生态进化学院∥生物防治国家重点实验室∥广东植物资源重点实验室,广东 广州510275
  • 收稿日期:2017-04-28 出版日期:2018-01-25 发布日期:2018-01-25
  • 通讯作者: 廖斌(1969年生),女;研究方向:超富集机理;E-mail:liaobin2005@126.com

The screening and expression pattern analysis of lead tolerant related gene in Viola baoshannensis

DENG Dongmei1, AI Hongxia2, DENG Jinchuan2, LIAO Bin2   

  1. 1.College of Biological and Chemical Engineering, Guangxi Key Laboratory of Green Processing of Sugar Resources, Guangxi University of Science and Technology,Liuzhou 545006,China;
    2. State Key Laboratory of Biocontrol, Guangdong Key Laboratory of Plant Resources, College of Ecology and Evolution, Sun Yat-sen University, Guangzhou 510275, China
  • Received:2017-04-28 Online:2018-01-25 Published:2018-01-25

摘要:

Cd超富集植物宝山堇菜Viola baoshannensis也有一定Pb耐性/富集能力,为了解宝山堇菜对Pb的耐性/富集分子机制,采用抑制消减杂交技术(suppression subtractive hybridization, SSH),筛选了宝山堇菜根在Pb诱导下差异表达的基因片段,进而对与Pb耐性与富集相关的基因克隆片段(expressed sequence tags, ESTs)进行半定量RT-PCR和Northorn杂交验证。利用SSH方法,从宝山堇菜根部获得27个在Pb胁迫下特异表达的无重复的有意义EST克隆片段。半定量RT-PCR和Northern杂交进一步证实和Lycopersicon esculentum木葡聚糖内源转糖基酶(xyloglucan endotransglycosylase, XTH)基因、Arabidopsis thaliana GTP结合蛋白基因(GTP binding mRNA)基因相似的2个克隆片段在宝山堇菜受Pb胁迫时,为上调表达基因。可初步假设,Pb胁迫下,伴随有细胞壁的变化,GTP 结合蛋白可能参与宝山堇菜体内Pb胁迫信号的传导。

关键词: 超富集植物, 宝山堇菜, Pb, SSH

Abstract:

Viola baoshanensis, an identified Cd hyperaccumulator can also tolerate and accumulate a certain amount of Pb. To understand the resistance/enrichment molecular mechanism of Viola baoshansis to Pb, suppression subtractive hybridization (SSH) was performed to screen gene segments induced by Pb, and semi-quantitive RT-PCR along with Northorn blotting was then used to verify the relationship between the screened expressed sequence tags (ESTs) and Pb tolerant/accumulation in V. baoshanensis. Twenty-seven different cDNA fragments were obtained via sequences analysis of cDNA inserts obtained from SSH subtractive cDNA library of V. baoshanensis roots. The results of both RT-PCR and Northern blot analysis showed that two cDNA fragments with high homology with xyloglucan endotransglycosylase (XTH) and GTP binding gene were up-regulated under Pb stress. It could be speculated that, in response to Pb stress, cell wall might have some changes and GTP binding protein might play a role in signal transduction.

Key words: hyperaccumualtor, Viola baoshanensis, Pb, SSH

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